10/19/2016

Biotechnology:

The Challenge of Tomorrow

Kamal Rashid, Ph.D. Director and Research Professor Worcester Polytechnic Institute

Biomanufacturing Education and Training Center Worcester Polytechnic Institute WPI -150 Years of Excellence in Higher Education

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Technology Progress

Chemistry & Physics First Half of 20th Century

Engineering, Electronics, Second Half of 20th Computer and Information Century Technology

Biotechnology, Bioprocessing, Dominant Economic Force Biomanufacturing, for the 21st Century Genomics, Proteomics and the rest of the “omics”

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Biotechnology • Putting biology to work for a better world

• The use of biological processes within the frame work of technical operations and industrial production: – Production of wine and beer – Production of cheese and bread – Production of biopharmaceutical

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Biotechnology: Historical Development

 Pre Pasteur Era Before 1865

 Alcoholic beverages, dairy products

 Pasteur Era 1865-1940

 Ethanol, acetone, glycerol, citric acid

 Antibiotic Era 1940-1960

 Penicillin, Viral vaccines

 Post Antibiotic Era 1960-1975

 Amino acids, enzymes

 Era of New Biotechnology 1975-

 Genetic Engineering

 Systems Biology

 CRISPR Technology- Exciting New Development

 Synthetic Biology

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Biotechnology Progress More than 250 healthcare products and vaccines

More than 50 biorefineries built in North America to produce: Biofuels Chemicals from renewable biomass

More than 13 million farmers around the globe user agricultural biotechnology to: Increase yield Control plant diseases Improve food quality

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Biotechnology Clusters

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FDA Product Approval 1996-2012

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U.S. Biopharma Pipeline

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Recombinant Organisms • Organisms which contain foreign genetic material, generally but not always extra chromosomal, that has been introduced deliberately as opposed to having been acquired naturally. • Genetically Engineered • Designed to carry out process efficiently – Overproduction of a protein not normally made by the host – Heavily utilized by the biotechnology industry

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Recombinant Proteins

Who are involved in the production ? Cell Biologists Chemical Engineers

• Gene Isolation *Large Scale production • Gene characterization *Optimize conditions • Gene modifications *Provide maximum yield • Create cells that effectively express genes • Use for industrial production of proteins

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Monoclonal Antibody Production

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So What is Bioprocessing ? Why use Biological Systems ? Biomanufacturing

Pharmaceuticals to Biopharmaceuticals

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Biomanufacturing

Cells Media Reactor

Conversion Stage DO pH Asepsis Agitation Aeration

Product Separation & Purification

Cell Disruption Centrifugation Filtration Chromatography Fill & Finish

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Bioprocessing Products • Biopharmaceuticals • Insulin • Growth hormones • Antibiotics • Vitamins • Biomass- Biofuels • Increased agricultural productivity – Food Bioprocessing • Lipases for cheese flavor and texture • Pectinases for clarification of wines and juices • Amylases for high Fructose corn syrup

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Bioprocessing Deals with Living Cells

• Microbial Cells • Animal Cells • Insect Cells • Plant Cells

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Microbial Cells Choice of the Organism • Most Common –Bacteria • E. Coli • Lactobacillus • Bacillus – Yeast • Saccharomyces •Pichia • Strain Improvement

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Microorganisms and Bioprocessing

• Ethanol production vs. amino acid production:

– Ethanol is a by product of catabolic pathways. These are waste products.

– Amino acids are building blocks that the cells synthesize at the expense of energy. This process is highly regulated.

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Microbial Metabolites • Primary metabolites: • Ethanol • Glutamic Acid & Citric Acid • Lysine • Polysaccharides • Secondary metabolites: – Produced mostly from intermediates and end products of primary metabolites: • Penicillin • Cephalosporin • Streptomycin • Cyclosporine

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Animal Cells and Bioprocessing

•BHK21 • Vero • HEK 293 • CHO • HeLa • 3T3Cells • Hundreds of other cell lines $1 invested in vaccine production saves $10 in future costs of health care

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Insect Cell Culture and Bioprocessing

Sf9 Cell Line Common name: Fall Armyworm

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Research Areas of Bioprocessing

• Bioreactor Design • Process Monitoring • Biocatalysis • Separation and Purification

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Biocatalysis

• Biocatalysis is the development of specialized enzymes and catalytic antibodies for particular uses. • A basic understanding of these biocatalysts at the molecular level is essential • Examples: – Thermodynamic studies to understand realizable yields. – Studies involving x-ray diffraction and nuclear magnetic resonance spectroscopy to elucidate atomic structure.

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Process Monitoring and Control

• Design of new and advanced methodologies for monitoring and controlling bioprocesses is essential . • Recent development in these methodologies include: – Capillary electrophoresis for rapid analytical techniques. – Designing of bio-sensing devices and control networks

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Separation and Purification

• There is a real need to improve the present methods of separating and purifying cellular products ( downstream processing). • Examples: – Development of industrial scale chromatographic separations. – Development of disposable industrial filtration technology – Cost effectiveness

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Cell Culture Process Development

Media Optimization Bioreactor Selection

Expression System

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Typical Biologic Process Flow

Cell Culture/Clarification

Cell Spinner or Production Depth Clarified bank Shake Seed Bioreactors Centrifugation vial Flask Bioreactor Filtration Harvest

Product Capture Fine Purification

Affinity low pH Viral Ion Exchange Hydrophobic Viral UF/DF Bottling Chromatography inactivation Chromatography Interaction Filtration Chromatography

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Agricultural Biotechnology Higher Quality Crops

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Agricultural Biotechnology Vaccines for Human Health Genetically engineered crops, usually fruits or vegetables, designed to carry antigenic proteins from infectious pathogens, that will trigger an immune response when ingested.

Tailor-made vaccines for cancer treatment have shown considerable promise in preliminary studies. Patient Specific.

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Agricultural Biotechnology More Products

Flowers: Improve Color, Smell and Size Biofuels: Provide the Feed stocks for Fermentation and Refining Animal and Plant Breeding: Marker Assisted Selection Pest Resistant Crops: Bt Corn Pesticide Resistant Crops: Selective Killing of Weeds Nutrient Supplementation: Golden Rice Industrial Strength Fiber: Spider Silk

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Agriculture Biotechnology Packaging to Reduce Spoilage

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Agricultural Biotechnology Reduced Health Risk

Genetically modified Vegetable Oils: More Linoleic Acid. The Beneficial Fatty Acid Reduced Saturated Fatty Acids

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Impact of Biotechnology on the Environment

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Biotechnology contributions to: Better understanding the environment Environmental restoration Environmental preservation

Biosystems for treatment of: Contaminated and radioactive waste Oil spills Acid wastes Municipal wastes Pesticides

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Prince William Sound- Alaska Remediation with Natural Bacteria

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Bioremediation

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Biosensors

 Biosensors help to know where and what the pollution is.  Identify the source and extent of the pollutants.  Monitor the site during and after the clean-up.  Give highly accurate results in short period of time.

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Functional Genomics

• Utilization of the massive amount od data generated by genome sequencing.

• Understanding the genome sequence is critical for the production of new biopharmaceutical products

• Advances in bioinformatics, high throughput technologies microarrays analysis and next generation DNA sequencing helped in advancing the field and identifying new and useful healthcare products

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What is Proteomics

The first use of the term “proteomics” was in 1995. Defined Then as the large-scale characterization of the entire protein complement of a cell line, tissue, or organism. Now two approaches exist: 1-Large scale analysis of gene products- Proteins 2-Comination of protein studies with genetics readout: -mRNA analysis -Genomics The goal of proteomics is: Obtain a more global and integrated view of biology by studying all the proteins of a cell rather than each one individually.

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Metabolomics and Bioprocessing • The era of OMICS • Metabolomics: ─ Identify, measure and interpret the complex time related concentration, activity and flux of endogenous metabolites in cells. • How metabolomics can impact biomanufacturing: ─ Improve yield ─ Enhance reproducibility ─ Increase process management ─ New- Started to be used in 2005. ─ Most of the published work has been on yeast and bacterial fermentation

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Systems Biology

• Moving away from local pathways to the whole system. • System biology: The study of systems of biological components, which may be molecules, cells, organisms or entire species. Living systems are dynamic and complex, and their behavior may be hard to predict from the properties of individual parts. • Systems biology is based on the understanding that the whole is greater than the sum of the parts.

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Systems Biology for Bioenergy

Source:U.S. Department of Energy Genomic Science program and provide the URL (http://genomicscience.energy.gov).

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DNA Repair and Cancer Cells

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CRISPR Technology

A new Genetic Engineering Technique What is CRISPR ? Clustered Regularly Interspaced Short Palindromic Repeat Faster and easier than other DNA modifying techniques It is set to revolutionize both basic research and disease treatment. These short repeats are found in the bacterial and other microorganisms genomes. They help in defending against viral invasions NIH has approved the first clinical trial using CRISPR

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MA Biopharma Job Market Listings

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Questions?

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For further information, please contact:

Dr. Kamal Rashid [email protected]

Director & Research Professor Worcester Polytechnic Institute Worcester, MA

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